Characterization of cultured synovial cells from human TMJ

本研究的目的是，首先，分离和培养滑膜细胞从人类颞下颌关节标本的特点，其次，调查这些细胞中的细胞因子和酶的生产。<Methods>采用体外培养法从TMJ滑膜组织中分离滑膜细胞，然后在含20%或10%胎牛血清的Ham F12中培养。使用免疫细胞化学检查滑膜细胞的细胞特异性标志物。接下来，用IL-1β或TNF-α处理滑膜细胞，IL-1β或TNF-α在TMJ疾病患者的滑液中增加。酶法测定与细胞外基质降解相关的纤溶酶原激活物（PA）活性。通过ELISA测量IL-6、IL-8和巨噬细胞引诱蛋白（MCP）-1的量。采用RTPCR方法研究了这些细胞因子的基因表达。<Rersults>滑膜细胞成纤维细胞特异性标记物和波形蛋白阳性。相反，细胞对巨噬细胞标志物、HLA II类和树突状细胞标志物呈阴性。然后，用或不用IL-1β处理滑膜细胞。IL-1β以时间和剂量依赖性方式增加滑膜细胞中IL-6、IL-8和MCP-1的产生。IL-1β刺激这些细胞因子的基因表达。IL-1β还增强滑膜细胞PA活性。然后用TNF-α处理细胞。TNF-α促进IL-8和MCP-1的产生和基因表达。<Conclusions>我们发现，培养的滑膜细胞可能为TMJ的细胞和分子机制的研究提供了重要的优势。此外，当TMJ疾病患者滑液中IL-1β和TNF-α水平升高时，滑膜细胞中PA活性和IL-6、IL-8和MCP-1的产生可能增加。

The aims of the present study were, firstly, to isolate and to characterize culture synovial cells obtained from human TMJ specimens, and secondly, to investigate the productions of cytokines and enzymes in those cells.<Methods> Synovial cells were isolated from TMJ synovial tissue using an out growth method and then cultured in Ham F12 containing 20% or 10% fetal bovine serum. Synovial cells were examined cell-specific markers using immunocytochemistry. In the next, synovial cells were treated with IL-1β or TNF-α, which was increased in synovial fluids from patients with TMJ disorders. Activity of plasminogen activator (PA), which is associated with extracellular matrix degradation, was measured by enzyme assay. Amounts of IL-6, IL-8 and macrophage attractant protein (MCP)-1 were measured by ELISA. Gene expressions of these cytokines were also investigated using RTPCR method.<Rersults> Synovial cells were positive for fibroblast-specific marker and vimentin. In contrast, the cells were negative for macrophage marker, HLA class II and dendritic cell marker. Then, synovial cells were treated with or without IL-1β. IL-1β increased IL-6, IL-8 and MCP-1 production in synovial cells in a time- and dose-dependent manner. IL-1β stimulated gene expression of these cytokines in them. IL-1β also enhanced PA activity in synovial cells. In the next, the cells were treated with TNF-α. TNF-α enhanced productions and gene expressions of IL-8 and MCP-1.<Conclusions> We found that cultured synovial cells might provide important advantages for studies of the cellular and molecular mechanisms in TMJ. In addition, PA activity and IL-6, IL-8, and MCP-1 productions may be increased in synovial cells when IL-1β and TNF-α levels are increased in synovial fluids from patients with TMJ disorders.